库尔勒香梨离体叶片再生体系的建立

【目的】建立库尔勒香梨叶片诱导再生的稳定高效再生体系。为梨的遗传转化研究奠定基础。【方法】以库尔勒香梨离体培养的继代30 d左右的无菌苗叶片为材料,1/2 MS为基本培养基,用 TDZ与 IBA或 NAA分别组合设计配方,研究不同生长调节剂质量浓度组合对不定芽诱导过程中愈伤组织形成率和不定芽形成率的影响,选取最优配方附加不同质量浓度的AgNO₃,获得不定芽再生的最佳培养基。用IBA和NAA分别组合设计生根生长调节剂的配比,根据生根率、平均生根数以及平均生根长度筛选最适宜生根的生长调节剂的配比,建立生根的培养体系。【结果】库尔勒香梨叶片诱导不定芽再生的最佳培养基为1/2 MS+TDZ 1.0 mg/L+IBA 0.5 mg/L+AgNO₃ 0.5 mg/L,愈伤组织诱导率100%,不定芽形成率84.92%。最适宜香梨再生苗生根的培养配方为1/2 MS + 0.5 mg/L IBA + 0.9 mg/L NAA,结合前期暗培养7 d,生根率可达100%,平均生根条数为5.66,平均根长度为 2.8 cm。【结论】建立了库尔勒香梨的再生体系。

Establishment of Regeneration System of Leaves in -vitro of Korla Fragrant Pear

【Objective】 The objective of the project is to establish the stable and effective regeneration system of leavesin -vitro of Korla Fragrant Pear seedlings.【Method】 The leaves of sterile seedlings of Korla fragrant pear cultured in vitro for about 30 days were used as the basic medium for 1/2 MS. The effects of different concentration of growth regulators on callus formation rate and adventitious bud formation rate during adventitious bud induction were studied by using TDZ (1.0 mg/L), IBA (0.3-0.9 mg/L) or NAA (0.3-0.9 mg/L) to design the formula separately to explore the effects of different concentration combinations of growth regulators on callus formation and adventitious bud formation during adventitious bud induction process. The optimal medium for adventitious bud regeneration was obtained by selecting the optimal formula and adding AgNo₃ with different mass concentration. The ratio of rooting growth regulators was designed with IBA and NAA respectively. According to the rooting rate, the average rooting number and the average rooting length, the ratio of the most suitable growth regulator were screened and the culture system of rooting was established.【Result】 The results indicated that the optimal medium for adventitious shoot induction was 1/2 MS supplemented with 1.0 mg/L TDZ, 0.5 mg/L IBA and 0.5 mg/L AgNO₃, the frequency of callus formation and adventitious shoot regeneration were 100 % and 84.92%, respectively. The appropriate rooting induction medium was 1/2 MS + 0.5 mg/L IBA + 0.9 mg/L NAA, the rooting rate could reach 100% after 7 days dark culture. The average rooting number of each stem was 5.66, and average rooting length was 2.8 cm.【Conclusion】 The regeneration system of leaves in -vitro of Korla Fragrant Pear seedlings was established, which laid foundation for genetic transformation of pears.